Mast Cell Development and Function in the Zebrafish

Abstract

The many advantages of the zebrafish model provide a unique opportunity to integrate the tools of developmental embryology, transgenesis, and functional assays to elucidate the molecular pathways underlying hematopoiesis and for modeling human blood diseases. These methodologies have recently been applied to the zebrafish mast cell lineage and have resulted in a better understanding of vertebrate mast cell biology. By employing whole-mount in situ hybridization alone and in combination with co-localization approaches, fluorescence-activated cell sorting (FACS), and morpholino gene knockdown studies, new insights into early mast cell transcriptional regulation and ontogeny have been exposed in vivo. Transgenic strategies have permitted the modeling of human mast cell diseases, like systemic mastocytosis in zebrafish, which can subsequently be exploited for high-throughput chemical screens to identify potential therapies in these conditions. Mast cell functional assays have been adapted to zebrafish providing the opportunity to utilize this model for interrogating the cellular players in innate and adaptive immunity and as a live animal readout for drug responses in allergic and inflammatory reactions. These techniques are detailed in the following chapter.

Sequential Immunofluorescence and Immunohistochemistry on Cryosectioned Zebrafish Embryos

“…We have presented a novel method for combined immunofluorescence and immunohistochemistry that represents an important step forward in performing colocalization experiments on cryosectioned zebrafish embryos. There is a critical lack of existing colocalization protocols that are optimized for use with small embryos and cryosectioned material 17,18 , both of which are otherwise commonly used in molecular studies . While these protocols can work well, their usefulness is limited by the availability of antibodies that (i) can be used in zebrafish and (ii) are compatible with IF.…”

“…Copyright © 2019 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License May 2019 | 147 | e59344 | Page 7 of 8Existing colocalization protocols mainly focus on simultaneous visualization of two fluorophores17,18 …”

Searching for tryptase in the RBL‐2H3 mast cell model: Preparation for comparative mast cell toxicology studies with zebrafish

“…Zebrafish ( Danio rerio ) (ZF) are a powerful, human‐disease relevant (Lieschke & Currie, ; Ward & Lieschke, ) model organism for toxicology studies (Garcia, Noyes, & Tanguay, ). ZF and humans share a highly conserved FcεRI receptor, and MCs of both species share similarities in innate and adaptive immune responses (Da’as et al, ; Da’as, Balci, & Berman, ; Dobson et al, ). Two methods for assessing MC function in ZF have recently been developed.…”

“…Da'as, Balci, & Berman, 2015;Dobson et al, 2008). Two methods for assessing MC function in ZF have recently been developed.…”

An Affordable Intraperitoneal Injection Setup for Juvenile and Adult Zebrafish

“…I ntraperitoneal (IP) injection of zebrafish is considered an effective and reproducible route of administration to study various biological 1,2 and chemical 3 agents. The tools that are currently used to perform IP injections in zebrafish have several limitations.…”